【作者】 仲山民；

【导师】 胡芳名；

【作者基本信息】 中南林学院 ， 森林培育， 2002， 博士

【摘要】 常山胡柚是中国柑桔属植物的一个新种，原产浙江省常山县。它自20世纪70年代末被发现后已多次在全国评比中获奖，并已成为我国的第一个优质杂柑、全国绿色食品、名特优果品等，是浙江省重点开发的、一个不可多得的“一优二高”经济树种。因此，对其进行深入细致、全面系统的研究就显得十分必要，也具有非常重要的现实意义及明显的社会和经济效益。国外至今尚无此种，也没有这方面的研究报道，所以对常山胡柚开展系列研究，还具有十分重要的学术价值。 本文在综述了常山胡柚研究进展的基础上，首先对其进行了分子生物学方面的研究探讨。经反复实验，最后采用改良的CTAB法，不仅成功地提取到常山胡柚叶片DNA，而且所得的DNA浓度高(平均为272.86μg·mL^-1，每克鲜叶片平均可得DNA约82μg)、纯度好（平均为1.56），完全符合RAPD分析的要求，也能基本满足其它分子生物学分析的要求。随后，又对常山胡柚目前所选并普遍推广应用的12个优良无性系及其它6个优株进行了RAPD分析。结果表明：实验中筛选确定的RAPD反应体系和扩增程序（条件）是完全适合的；所筛选出的12个随机引物共得到随机扩增位点103个，平均每个引物产生8.6个识别位点；但其中均未发现多态性位点。由此便可大致推断：目前所选的12个常山胡柚优良无性系及其它6个优株之间在分子水平上并没有什么差异，其经济性状方面表现出的差异可能主要受生长发育阶段、生态环境条件及经营管理水平等因素的影响。 在浙江省常山县设立试验地，分别于2000～2001年采用标准株法，对其不同方位的果实进行定期定果的测定，然后进行统计分析，结果表明：常山胡柚果实在生长发育过程中，其大小的变化规律是随着生长发育时间的延长而不断的增长，其中有两个明显的增长快速期，分别处于7月的上半个月和9月的上中旬；其重量的变化规律也是随着生长发育时间的延长而不断的增长，但是，它与果实大小的增长正好相错。果实纵径与横径的变化规律非常一致，只是横径的增长速度明显快于纵径的增长速度，特别是在8月上中旬时表现尤为突出。不同的方位对果实的生长发育有一定的影响，方差分析也表明：果实的纵径与横径，在不同的方位之间、在不同的生长发育时期均存在着极显著的差异；而果形指数、果实重量的变化，在不同方位之间都没有显著性差异，但在不同的生长发育时期却有着极显著的差异。常山胡柚果实成熟后，不论大小如何，其果形指数非常相近，均在0.87左右，表明它们的果形差异不明显，难以作为区分类型的依据。 采用常规分析法对常山胡柚3种不同大小类型的果实及目前普遍推广应用的12个优良无性系果实的品质及其在贮藏期间的变化情况进行了分析与统计，结果表明：常山胡柚各类型、各无性系的果实在贮藏期间，其重量不断减轻，可食部分比例不断增加；各部位的含水量总体呈下降趋势，其中果皮的含水量下降最大，其次为白皮层和囊衣，而果肉中的变幅最小，且始终保持在87～92％的高水平上，可见，常山胡柚果实的贮藏保鲜能力很强；其含酸量总体呈下降趋势；可溶性固形物含量变化起伏较大，而总糖含量和还原糖含量的变化趋势 学位论文：摘要较一致，先升后降又逐渐上升；果实的糖酸比值在不断提高，致使风味越来越好；果实中VC含量的变化规律性不明显，但总体略有上升；果皮中SOD活性的变化规律是先升后降又明显上升，并保持在较高水平上，而果肉中的SOD活性则一直保持平稳而缓慢的上升，这可能就是常山胡袖果实之所以特别耐藏的原因之一；果实中的POD活性主要存在于果皮组织中，且呈波浪状变化：其＊AT活性的变化起伏也较大，尤其是果皮组织中的＊AT活性波动更为明显；果皮和果肉的细胞膜相对透性均呈上升趋势，其中果皮的上升幅度达59．8％，果肉的上升幅度仅为门石％，表明果皮的衰老明显快于果肉。上述这些变化，在不同的类型、不同的忧良无性系之间均表现出程度不一的差异显著性。 对常山胡抽果实进行加工利用的必要性、依据、途径及其现状作了充分的论述，并先后开展了常山胡袖果汁饮料、低糖果酱、砂囊悬浮果汁、砂囊罐头、果脯等系列产品的加工工艺及其关键技术（如果汁脱苦、果汁稳定性、砂囊悬浮效果、各产品品质等）的研究，结果显示：对常山胡抽鲜果通过适当的处理，将其制成风味独特且富有营养的系列产品完全可行；常山胡抽鲜果不论大小如何，其出汁率都在50％以上，平均为52二％，而且3种不同大小类型果实在出汁率方面并无显著性差异；常山胡柏果肉浆液在低于70C的温度条件下，并不表现苦味，当温度上升到70oC以后，随着加热温度的提高、加热时间的延长，其苦味逐渐呈现并加重，当瞬间快速加热到某一温度时，常山胡抽果肉浆液的苦味变化不大，随着在某一温度下维持时间的延长，其苦味则不断加重。加工中可以通过添加 03～0．5％比例的D一CD来避免或减轻常山胡柏果汁产品的苦味；在常山胡袖果汁及其饮料加工中，进行高压均质处理后，其稳定性明显提高，而且以采用 40 MPa的压力对常山胡抽果汁处理 1次或采用30 MPa的压力对其处理2次，更多还原

【Abstract】 Changshan-huyou is a new species of citrus native to Changshan County, Zhejiang Province. Since its discovery at the end of 1970s, it has won several prizes in different national appraisals. As the first top-quality hybrid citrus in China, national green food and brand-name and quality fruit, Changshan-huyou is a rare major developed top-quality high-efficient and high production economic tree species. Hence, it is necessary to make in-depth and all-round systematic researches on Changshan-huyou. The researches are of vital realistic significance and distinct social and economic efficiency. There is no report on such researches abroad, so these series of researches will even be of very important academic value.This paper makes a molecular biological study on Changshan-huyou on the basis of summarizing the progress of researches on it. After repeated experiments, through adopting the improved CTAB method, we succeed in extracting high-density and purity DNA from the leaf of Changshan-huyou. The density averages 272.86#g ?mL-1, and about 82#g DNA can be obtained from per gram’s fresh leaf. The purity reaches 1.56 on average. Both of the density and purity fully meet the demands of RAPD analysis and other molecular biological analysis. The paper also makes RAPD analysis of 12 popularized elite clones and 6 other fine plants. It proves that the RAPD response system determined in the experiment fully fits the cloning procedure(condition); 103 random cloning sites are achieved from 12 random chosen primers. On average, every primer creates 8.6 identifying sites, yet no polymorphic site is discovered. It infers from the analysis that there is no molecular difference between the 12 chosen elite clones of Changshan-huyou and the 6 other fine plants . The economic quality difference is mainly influenced by such factors as the phase of growth, the condition of ecological environment and the level of management.At the experiment site in Changshan county of Zhejiang province, standard trees were selected in 2000 and 2001. The fruits at different orientation were examined and analyzed. The result shows that the size of fruit increases in shaped form with the prolongation of growth. There are two rapid growth periods in a year: the early part of July and the mid September. The weight increases likewise but it alternates with the increase of fruit size. The change of the vertical diameter and the horizontal diameter are very similar except that the latter increases obviously faster than the vertical diameter does, especially in early August. The orientation has effect on the growth to some extent. It is showed by the standard deviation analysis that the vertical and the horizontal diameter have significant difference in different orientations and growth periods. The index of fruit and the change of the weight of fruits have no significant difference in various orientations, while they have in different growth periods. The fruits of Changshan-huyou have very similar index as 0.87 when they are ripe, which shows that they can not be classified by index since there is no obvious difference in the shape of fruits.By means of routine analysis, the quality and the variation during the storage of fruits of 3 different types of Changshan-huyou and 12 elite clones are analyzed and counted. The findings show that the weight of various types and clones of Changshan-huyou decrease in the course of storage, while the edible ratio increases continuously. The water content in all parts is on decrease, of which the greatest decrease is in that of the pericarp. The decrease in the white seedcase and thecapsular cover follow next. But the water content in sarcocarp changes very little and keeps at the high level of 87-92%. Therefore, there is an evidence that the fruit of Changshan-huyou has a strong capability of retain freshness. The acid content decreases and the content of soluble solid fluctuate greatly. The content of total sugar and reducing sugar are similar in change, appearing an up-down- up trend. The ratios更多还原